11836170001 superasin thermo am2696 rnasin promega n2515  (Thermo Fisher)


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    Name:
    SUPERase• In RNase Inhibitor 20 U μL
    Description:
    SUPERase• In RNase Inhibitor is a protein based inhibitor of nonhuman origin that noncovalently binds and inhibits the most common and troublesome RNases including RNase A B C 1 and T1 Features of SUPERase• In RNase Inhibitor • Completely removes RNase contamination from glass and plastic surfaces• Excels at removing high levels of RNase contamination where similar products fail• Proven effective at removing high concentrations of dried on RNase A• Ideal for cleaning work surfaces pipettors and equipment that must be RNase freeSUPERase• In RNase Inhibitor can be used in any application where RNase contamination could be problematic Because it inhibits a broader range of RNases than traditional RNase inhibitors SUPERase• In is the most effective RNase inhibitor available providing a higher level of protection against degradation SUPERase• In does not interfere with other enzymes such as RNA polymerases reverse transcriptase or Taq DNA polymerase Additionally SUPERase• In is active up to 65°C and over a pH range of 5 5 to 8 5 If you are plannning to use this product with standard antibody purification methods please contact our technical support to discuss experimental design Unit definition SUPERase• In RNase Inhibitor at 1 U µL will block the degradation of 0 1 µg µL labeled RNA by 2 5 pg µL of RNase A 2 5 pg µL of RNase I and 0 0075 U µL of RNase T1 for 4 hours at 37°C in 20 mM Tris HCl pH 7 5 50 mM NaCl 1 mM EDTA Analysis is by denaturing PAGE SUPERase• In is currently the only ribonuclease inhibitor for which the unit activity is defined by such a functional assay
    Catalog Number:
    am2694
    Price:
    None
    Applications:
    General Real-Time PCR Reagents|In Vitro Transcription|PCR & Real-Time PCR|Real Time PCR (qPCR)|Reverse Transcription|Gene Expression Analysis & Genotyping
    Category:
    Proteins Enzymes Peptides
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    Structured Review

    Thermo Fisher 11836170001 superasin thermo am2696 rnasin promega n2515
    SUPERase• In RNase Inhibitor is a protein based inhibitor of nonhuman origin that noncovalently binds and inhibits the most common and troublesome RNases including RNase A B C 1 and T1 Features of SUPERase• In RNase Inhibitor • Completely removes RNase contamination from glass and plastic surfaces• Excels at removing high levels of RNase contamination where similar products fail• Proven effective at removing high concentrations of dried on RNase A• Ideal for cleaning work surfaces pipettors and equipment that must be RNase freeSUPERase• In RNase Inhibitor can be used in any application where RNase contamination could be problematic Because it inhibits a broader range of RNases than traditional RNase inhibitors SUPERase• In is the most effective RNase inhibitor available providing a higher level of protection against degradation SUPERase• In does not interfere with other enzymes such as RNA polymerases reverse transcriptase or Taq DNA polymerase Additionally SUPERase• In is active up to 65°C and over a pH range of 5 5 to 8 5 If you are plannning to use this product with standard antibody purification methods please contact our technical support to discuss experimental design Unit definition SUPERase• In RNase Inhibitor at 1 U µL will block the degradation of 0 1 µg µL labeled RNA by 2 5 pg µL of RNase A 2 5 pg µL of RNase I and 0 0075 U µL of RNase T1 for 4 hours at 37°C in 20 mM Tris HCl pH 7 5 50 mM NaCl 1 mM EDTA Analysis is by denaturing PAGE SUPERase• In is currently the only ribonuclease inhibitor for which the unit activity is defined by such a functional assay
    https://www.bioz.com/result/11836170001 superasin thermo am2696 rnasin promega n2515/product/Thermo Fisher
    Average 99 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    11836170001 superasin thermo am2696 rnasin promega n2515 - by Bioz Stars, 2020-04
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Translation Enhancing ACA Motifs and Their Silencing by a Bacterial Small Regulatory RNA
    Article Snippet: Genes to be analyzed were cloned under T7 promoter control in the DFRH plasmid provided with the kit. .. In addition to kit solutions A and B, reaction mix contained, 10 U of RNase inhibitor SUPERase (Ambion) and template plasmid DNA added to either 0.5 or 5 pM final concentration.

    Centrifugation:

    Article Title: Enhanced Replication of Mouse Adenovirus Type 1 following Virus-Induced Degradation of Protein Kinase R (PKR)
    Article Snippet: Cells were collected at 24 hpi by scraping in ice-cold PBS containing 100 µg/ml cycloheximide, pelleting, and resuspending in 500 µl lysis buffer (20 mM Tris-Cl, 150 mM NaCl, 15 mM MgCl2 , 8% glycerol, 20 IU/ml SUPERase•In [Thermo Fisher Scientific catalog no. AM2696], 80 IU/ml murine RNase inhibitor [New England BioLabs catalog no. M0314S], 0.1 mg/ml heparin [Sigma H3393-50], 0.1 mg/ml cycloheximide, 1 mM DTT, 1× protease inhibitor [protease inhibitor cocktail kit; Thermo Scientific catalog no. 78410], 20 IU/ml Turbo DNase [Thermo Fisher Scientific catalog no. AM2238], 1% Triton X-100 [Sigma T9284]). .. Cells were lysed by passage through a chilled 26-gauge needle 10 times, vortex mixing for 30 s, and then incubating on ice for 5 min. Lysates were cleared by centrifugation for 5 min at 14,000 × g at 4°C.

    Article Title: Transcriptional response to stress is pre-wired by promoter and enhancer architecture
    Article Snippet: After centrifugation (1000 g, 5 min, 4 °C), the nuclei were collected and flash frozen in storage buffer (10 mM Tris-HCl pH 8.0, 25% glycerol, 5 mM MgAc2 , 0.1 mM EDTA, 5 mM DTT). .. The nuclear run-on reactions were performed at 37 °C for 3 min in the presence of 0.05 mM biotin-A/C/G/UTP (Perkin Elmer), 0.5% sarkosyl, 5 mM Tris-HCl (pH 8.0), 1.5 mM MgCl2 , 0.5 mM DTT, 150 mM KCl and 0.1 units per ul Superase RNase inhibitor (Life Technologies).

    Amplification:

    Article Title: KSHV induces immunoglobulin rearrangements in mature B lymphocytes
    Article Snippet: Single cell RT-PCR for immunoglobulin light chains Single cells were harvested by flow sorting into 96-well PCR plates containing 4μl of RNA lysis buffer (0.5x PBS+10mM DTT+4U SUPERas-In (Thermo Cat #AM2694)). .. PCR primers for amplification of immunoglobulin light chains were from Tiller et. al.[ ].

    Article Title: KSHV induces immunoglobulin rearrangements in mature B lymphocytes
    Article Snippet: Single cells were harvested by flow sorting into 96-well PCR plates containing 4μl of RNA lysis buffer (0.5x PBS+10mM DTT+4U SUPERas-In (Thermo Cat #AM2694)). .. PCR primers for amplification of immunoglobulin light chains were from Tiller et. al.[ ].

    Synthesized:

    Article Title: KSHV induces immunoglobulin rearrangements in mature B lymphocytes
    Article Snippet: Single cell RT-PCR for immunoglobulin light chains Single cells were harvested by flow sorting into 96-well PCR plates containing 4μl of RNA lysis buffer (0.5x PBS+10mM DTT+4U SUPERas-In (Thermo Cat #AM2694)). .. Plates were sealed and stored at -80°C. cDNA was synthesized directly in wells using Thermo High Capacity cDNA synthesis kit (Cat #4368814).

    Article Title: KSHV induces immunoglobulin rearrangements in mature B lymphocytes
    Article Snippet: Single cells were harvested by flow sorting into 96-well PCR plates containing 4μl of RNA lysis buffer (0.5x PBS+10mM DTT+4U SUPERas-In (Thermo Cat #AM2694)). .. Plates were sealed and stored at -80°C. cDNA was synthesized directly in wells using Thermo High Capacity cDNA synthesis kit (Cat #4368814).

    Construct:

    Article Title: Unique Features of the m6A Methylome in Arabidopsis thaliana
    Article Snippet: The eluted RNA was treated with RNasin (Ambion Cat No. AM2694) according to the manufacturer’s instructions. .. TruSeq Stranded mRNA Sample Prep Kit (Illumina) was used to construct the library from immunoprecipitated RNA and input RNA according to a published protocol .

    Real-time Polymerase Chain Reaction:

    Article Title: Accurate, Streamlined Analysis of mRNA Translation by Sucrose Gradient Fractionation
    Article Snippet: Reagents Cells-to-Ct kit (Thermo Fisher Scientific, Invitrogen™, catalog number: AM1728) Superase-In (Thermo Fisher Scientific, Invitrogen™, catalog number: AM2696) XenoRNA (Thermo Fisher Scientific, Invitrogen™, catalog number: 4386995, part of control kit) Note: Store in small aliquots at −80 °C. .. TaqMan Gene Expression Master mix (Thermo Fisher Scientific, Applied Biosystems™, catalog number: 4369016) Primer/Probe qPCR assays for genes of interest (Thermo Fisher Scientific, Ac00010014_a1 (XenoRNA) and Mr03987587_mr (Luciferase))

    Incubation:

    Article Title: RNA-Guided RNA Cleavage by a CRISPR RNA-Cas Protein Complex
    Article Snippet: The resulting RNA products were isolated by PCI extraction and ethanol precipitation, and subject to polyadenylation by incubation with 5 U E. coli polyA polymerase (NEB) for 15 minutes at 37°C as described by the manufacturer. .. In order to determine the divalent metal requirements of the purified complex, cleavage reactions were performed for 1 hour at 70°C in 50 mM HEPES pH 7.0, 250 mM KCl, 1 mM ATP, 10 mM DTT, 0.1 mM EDTA, and 1 mM metal (if applicable) in the presence of 1 unit of SUPERase-In ribonuclease inhibitor (Applied Biosystems).

    Article Title: Transcriptional response to stress is pre-wired by promoter and enhancer architecture
    Article Snippet: In detail, the untreated or heat-treated samples were washed with PBS, and incubated for 5 min in nuclear isolation buffer (10 mM Tris-Cl pH 8.0, 300 mM sucrose, 3 mM CaCl2 , 2 mM MgAc2 , 0.1% TritonX-100, 0.5 mM DTT), followed by 25x douncing (Wheaton, #357546, loose pestle), to isolate the nuclei. .. The nuclear run-on reactions were performed at 37 °C for 3 min in the presence of 0.05 mM biotin-A/C/G/UTP (Perkin Elmer), 0.5% sarkosyl, 5 mM Tris-HCl (pH 8.0), 1.5 mM MgCl2 , 0.5 mM DTT, 150 mM KCl and 0.1 units per ul Superase RNase inhibitor (Life Technologies).

    Article Title: Unique Features of the m6A Methylome in Arabidopsis thaliana
    Article Snippet: The mixture was then incubated with protein-A beads and eluted with elution buffer (1× IP buffer and 6.7 mM m6 A). .. The eluted RNA was treated with RNasin (Ambion Cat No. AM2694) according to the manufacturer’s instructions.

    Luciferase:

    Article Title: Accurate, Streamlined Analysis of mRNA Translation by Sucrose Gradient Fractionation
    Article Snippet: Reagents Cells-to-Ct kit (Thermo Fisher Scientific, Invitrogen™, catalog number: AM1728) Superase-In (Thermo Fisher Scientific, Invitrogen™, catalog number: AM2696) XenoRNA (Thermo Fisher Scientific, Invitrogen™, catalog number: 4386995, part of control kit) Note: Store in small aliquots at −80 °C. .. TaqMan Gene Expression Master mix (Thermo Fisher Scientific, Applied Biosystems™, catalog number: 4369016) Primer/Probe qPCR assays for genes of interest (Thermo Fisher Scientific, Ac00010014_a1 (XenoRNA) and Mr03987587_mr (Luciferase))

    Expressing:

    Article Title: Accurate, Streamlined Analysis of mRNA Translation by Sucrose Gradient Fractionation
    Article Snippet: Reagents Cells-to-Ct kit (Thermo Fisher Scientific, Invitrogen™, catalog number: AM1728) Superase-In (Thermo Fisher Scientific, Invitrogen™, catalog number: AM2696) XenoRNA (Thermo Fisher Scientific, Invitrogen™, catalog number: 4386995, part of control kit) Note: Store in small aliquots at −80 °C. .. TaqMan Gene Expression Master mix (Thermo Fisher Scientific, Applied Biosystems™, catalog number: 4369016) Primer/Probe qPCR assays for genes of interest (Thermo Fisher Scientific, Ac00010014_a1 (XenoRNA) and Mr03987587_mr (Luciferase))

    Western Blot:

    Article Title: Translation Enhancing ACA Motifs and Their Silencing by a Bacterial Small Regulatory RNA
    Article Snippet: In addition to kit solutions A and B, reaction mix contained, 10 U of RNase inhibitor SUPERase (Ambion) and template plasmid DNA added to either 0.5 or 5 pM final concentration. .. Aliquots were loaded on 12.5% Acrylamide gels and Western analysis performed as previously described .

    Flow Cytometry:

    Article Title: KSHV induces immunoglobulin rearrangements in mature B lymphocytes
    Article Snippet: .. Single cell RT-PCR for immunoglobulin light chains Single cells were harvested by flow sorting into 96-well PCR plates containing 4μl of RNA lysis buffer (0.5x PBS+10mM DTT+4U SUPERas-In (Thermo Cat #AM2694)). .. Plates were sealed and stored at -80°C. cDNA was synthesized directly in wells using Thermo High Capacity cDNA synthesis kit (Cat #4368814).

    Article Title: KSHV induces immunoglobulin rearrangements in mature B lymphocytes
    Article Snippet: .. Single cells were harvested by flow sorting into 96-well PCR plates containing 4μl of RNA lysis buffer (0.5x PBS+10mM DTT+4U SUPERas-In (Thermo Cat #AM2694)). .. Plates were sealed and stored at -80°C. cDNA was synthesized directly in wells using Thermo High Capacity cDNA synthesis kit (Cat #4368814).

    Concentration Assay:

    Article Title: RNA-Guided RNA Cleavage by a CRISPR RNA-Cas Protein Complex
    Article Snippet: In order to determine the divalent metal requirements of the purified complex, cleavage reactions were performed for 1 hour at 70°C in 50 mM HEPES pH 7.0, 250 mM KCl, 1 mM ATP, 10 mM DTT, 0.1 mM EDTA, and 1 mM metal (if applicable) in the presence of 1 unit of SUPERase-In ribonuclease inhibitor (Applied Biosystems). .. Certified metal reference solutions (Spex CertiPrep except calcium obtained from Fisher Scientific) were added to 1 mM final concentration.

    Article Title: Translation Enhancing ACA Motifs and Their Silencing by a Bacterial Small Regulatory RNA
    Article Snippet: .. In addition to kit solutions A and B, reaction mix contained, 10 U of RNase inhibitor SUPERase (Ambion) and template plasmid DNA added to either 0.5 or 5 pM final concentration. ..

    Protease Inhibitor:

    Article Title: Enhanced Replication of Mouse Adenovirus Type 1 following Virus-Induced Degradation of Protein Kinase R (PKR)
    Article Snippet: .. Cells were collected at 24 hpi by scraping in ice-cold PBS containing 100 µg/ml cycloheximide, pelleting, and resuspending in 500 µl lysis buffer (20 mM Tris-Cl, 150 mM NaCl, 15 mM MgCl2 , 8% glycerol, 20 IU/ml SUPERase•In [Thermo Fisher Scientific catalog no. AM2696], 80 IU/ml murine RNase inhibitor [New England BioLabs catalog no. M0314S], 0.1 mg/ml heparin [Sigma H3393-50], 0.1 mg/ml cycloheximide, 1 mM DTT, 1× protease inhibitor [protease inhibitor cocktail kit; Thermo Scientific catalog no. 78410], 20 IU/ml Turbo DNase [Thermo Fisher Scientific catalog no. AM2238], 1% Triton X-100 [Sigma T9284]). .. Cells were lysed by passage through a chilled 26-gauge needle 10 times, vortex mixing for 30 s, and then incubating on ice for 5 min. Lysates were cleared by centrifugation for 5 min at 14,000 × g at 4°C.

    Infection:

    Article Title: Enhanced Replication of Mouse Adenovirus Type 1 following Virus-Induced Degradation of Protein Kinase R (PKR)
    Article Snippet: The next day, the cells were infected with MAV-1 at an MOI of 2. .. Cells were collected at 24 hpi by scraping in ice-cold PBS containing 100 µg/ml cycloheximide, pelleting, and resuspending in 500 µl lysis buffer (20 mM Tris-Cl, 150 mM NaCl, 15 mM MgCl2 , 8% glycerol, 20 IU/ml SUPERase•In [Thermo Fisher Scientific catalog no. AM2696], 80 IU/ml murine RNase inhibitor [New England BioLabs catalog no. M0314S], 0.1 mg/ml heparin [Sigma H3393-50], 0.1 mg/ml cycloheximide, 1 mM DTT, 1× protease inhibitor [protease inhibitor cocktail kit; Thermo Scientific catalog no. 78410], 20 IU/ml Turbo DNase [Thermo Fisher Scientific catalog no. AM2238], 1% Triton X-100 [Sigma T9284]).

    Polymerase Chain Reaction:

    Article Title: Diverse motif ensembles specify non-redundant DNA binding activities of AP-1 family members in macrophages
    Article Snippet: Collect beads on magnet and transfer eluate containing fragmented mRNA to a new PCR strip. .. First strand synthesis: We mixed fragmented RNA with 0.5 μL random primer (3 μg/μL) Life Tech #48190-011, 0.5 μL oligo-dT (50 μM from SSIII kit), 1 μL dNTPs (10 mM Life Tech, cat 18427088) and 0.5 μL SUPERase-In (ThermoFisher Cat#AM2696) and heat 50 °C for 1 min.

    Article Title: KSHV induces immunoglobulin rearrangements in mature B lymphocytes
    Article Snippet: .. Single cell RT-PCR for immunoglobulin light chains Single cells were harvested by flow sorting into 96-well PCR plates containing 4μl of RNA lysis buffer (0.5x PBS+10mM DTT+4U SUPERas-In (Thermo Cat #AM2694)). .. Plates were sealed and stored at -80°C. cDNA was synthesized directly in wells using Thermo High Capacity cDNA synthesis kit (Cat #4368814).

    Article Title: KSHV induces immunoglobulin rearrangements in mature B lymphocytes
    Article Snippet: .. Single cells were harvested by flow sorting into 96-well PCR plates containing 4μl of RNA lysis buffer (0.5x PBS+10mM DTT+4U SUPERas-In (Thermo Cat #AM2694)). .. Plates were sealed and stored at -80°C. cDNA was synthesized directly in wells using Thermo High Capacity cDNA synthesis kit (Cat #4368814).

    Article Title: Desmoplasia and oncogene driven acinar-to-ductal metaplasia are concurrent events during acinar cell-derived pancreatic cancer initiation in young adult mice
    Article Snippet: RNA was eluted using 11 μL of RNase Free ddH2 O and 0.5 μL of SUPERase•In RNase Inhibitor (Thermo Fisher, Cat# AM2694) was immediately added to eluted RNA. .. A 1 μL aliquot of each RNA sample was then stored into a separate 0.2 μL Axygen PCR tube (Thermo Fisher, Cat# 14-222-261) and this sample was used for RNA QC.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: KSHV induces immunoglobulin rearrangements in mature B lymphocytes
    Article Snippet: .. Single cell RT-PCR for immunoglobulin light chains Single cells were harvested by flow sorting into 96-well PCR plates containing 4μl of RNA lysis buffer (0.5x PBS+10mM DTT+4U SUPERas-In (Thermo Cat #AM2694)). .. Plates were sealed and stored at -80°C. cDNA was synthesized directly in wells using Thermo High Capacity cDNA synthesis kit (Cat #4368814).

    Article Title: KSHV induces immunoglobulin rearrangements in mature B lymphocytes
    Article Snippet: Paragraph title: Single cell RT-PCR for immunoglobulin light chains ... Single cells were harvested by flow sorting into 96-well PCR plates containing 4μl of RNA lysis buffer (0.5x PBS+10mM DTT+4U SUPERas-In (Thermo Cat #AM2694)).

    Magnetic Beads:

    Article Title: Transcriptional response to stress is pre-wired by promoter and enhancer architecture
    Article Snippet: The nuclear run-on reactions were performed at 37 °C for 3 min in the presence of 0.05 mM biotin-A/C/G/UTP (Perkin Elmer), 0.5% sarkosyl, 5 mM Tris-HCl (pH 8.0), 1.5 mM MgCl2 , 0.5 mM DTT, 150 mM KCl and 0.1 units per ul Superase RNase inhibitor (Life Technologies). .. For high specificity, the biotinylated nascent transcripts were purified three times with streptavidin-coated magnetic beads (Life Technologies), each round followed by isolation with Trizol (Life Technlogies) and EtOH-precipitation.

    Mutagenesis:

    Article Title: Translation Enhancing ACA Motifs and Their Silencing by a Bacterial Small Regulatory RNA
    Article Snippet: The hybrid genes carried yifK wt or mutant 5′ UTR sequences fused to the cat -3×FLAG coding sequence (chloramphenicol acetyl transferase in-frame fusion to the 3×FLAG epitope). .. In addition to kit solutions A and B, reaction mix contained, 10 U of RNase inhibitor SUPERase (Ambion) and template plasmid DNA added to either 0.5 or 5 pM final concentration.

    Isolation:

    Article Title: RNA-Guided RNA Cleavage by a CRISPR RNA-Cas Protein Complex
    Article Snippet: The resulting RNA products were isolated by PCI extraction and ethanol precipitation, and subject to polyadenylation by incubation with 5 U E. coli polyA polymerase (NEB) for 15 minutes at 37°C as described by the manufacturer. .. In order to determine the divalent metal requirements of the purified complex, cleavage reactions were performed for 1 hour at 70°C in 50 mM HEPES pH 7.0, 250 mM KCl, 1 mM ATP, 10 mM DTT, 0.1 mM EDTA, and 1 mM metal (if applicable) in the presence of 1 unit of SUPERase-In ribonuclease inhibitor (Applied Biosystems).

    Article Title: Diverse motif ensembles specify non-redundant DNA binding activities of AP-1 family members in macrophages
    Article Snippet: Paragraph title: PolyA RNA isolation and fragmentation ... First strand synthesis: We mixed fragmented RNA with 0.5 μL random primer (3 μg/μL) Life Tech #48190-011, 0.5 μL oligo-dT (50 μM from SSIII kit), 1 μL dNTPs (10 mM Life Tech, cat 18427088) and 0.5 μL SUPERase-In (ThermoFisher Cat#AM2696) and heat 50 °C for 1 min.

    Article Title: Transcriptional response to stress is pre-wired by promoter and enhancer architecture
    Article Snippet: In detail, the untreated or heat-treated samples were washed with PBS, and incubated for 5 min in nuclear isolation buffer (10 mM Tris-Cl pH 8.0, 300 mM sucrose, 3 mM CaCl2 , 2 mM MgAc2 , 0.1% TritonX-100, 0.5 mM DTT), followed by 25x douncing (Wheaton, #357546, loose pestle), to isolate the nuclei. .. The nuclear run-on reactions were performed at 37 °C for 3 min in the presence of 0.05 mM biotin-A/C/G/UTP (Perkin Elmer), 0.5% sarkosyl, 5 mM Tris-HCl (pH 8.0), 1.5 mM MgCl2 , 0.5 mM DTT, 150 mM KCl and 0.1 units per ul Superase RNase inhibitor (Life Technologies).

    Article Title: Unique Features of the m6A Methylome in Arabidopsis thaliana
    Article Snippet: Polyadenylated RNA was extracted using FastTrack MAG Maxi mRNA isolation kit (Invitrogen). .. The eluted RNA was treated with RNasin (Ambion Cat No. AM2694) according to the manufacturer’s instructions.

    Article Title: Desmoplasia and oncogene driven acinar-to-ductal metaplasia are concurrent events during acinar cell-derived pancreatic cancer initiation in young adult mice
    Article Snippet: Paragraph title: RNA isolation ... RNA was eluted using 11 μL of RNase Free ddH2 O and 0.5 μL of SUPERase•In RNase Inhibitor (Thermo Fisher, Cat# AM2694) was immediately added to eluted RNA.

    Labeling:

    Article Title: RNA-Guided RNA Cleavage by a CRISPR RNA-Cas Protein Complex
    Article Snippet: For analysis of the chemical ends of the cleavage products, cleavage reactions were performed using 5’-end labeled target as described above. .. In order to determine the divalent metal requirements of the purified complex, cleavage reactions were performed for 1 hour at 70°C in 50 mM HEPES pH 7.0, 250 mM KCl, 1 mM ATP, 10 mM DTT, 0.1 mM EDTA, and 1 mM metal (if applicable) in the presence of 1 unit of SUPERase-In ribonuclease inhibitor (Applied Biosystems).

    Purification:

    Article Title: RNA-Guided RNA Cleavage by a CRISPR RNA-Cas Protein Complex
    Article Snippet: .. In order to determine the divalent metal requirements of the purified complex, cleavage reactions were performed for 1 hour at 70°C in 50 mM HEPES pH 7.0, 250 mM KCl, 1 mM ATP, 10 mM DTT, 0.1 mM EDTA, and 1 mM metal (if applicable) in the presence of 1 unit of SUPERase-In ribonuclease inhibitor (Applied Biosystems). .. Certified metal reference solutions (Spex CertiPrep except calcium obtained from Fisher Scientific) were added to 1 mM final concentration.

    Article Title: Transcriptional response to stress is pre-wired by promoter and enhancer architecture
    Article Snippet: The nuclear run-on reactions were performed at 37 °C for 3 min in the presence of 0.05 mM biotin-A/C/G/UTP (Perkin Elmer), 0.5% sarkosyl, 5 mM Tris-HCl (pH 8.0), 1.5 mM MgCl2 , 0.5 mM DTT, 150 mM KCl and 0.1 units per ul Superase RNase inhibitor (Life Technologies). .. For high specificity, the biotinylated nascent transcripts were purified three times with streptavidin-coated magnetic beads (Life Technologies), each round followed by isolation with Trizol (Life Technlogies) and EtOH-precipitation.

    Sequencing:

    Article Title: Translation Enhancing ACA Motifs and Their Silencing by a Bacterial Small Regulatory RNA
    Article Snippet: The hybrid genes carried yifK wt or mutant 5′ UTR sequences fused to the cat -3×FLAG coding sequence (chloramphenicol acetyl transferase in-frame fusion to the 3×FLAG epitope). .. In addition to kit solutions A and B, reaction mix contained, 10 U of RNase inhibitor SUPERase (Ambion) and template plasmid DNA added to either 0.5 or 5 pM final concentration.

    Article Title: Unique Features of the m6A Methylome in Arabidopsis thaliana
    Article Snippet: Paragraph title: High-throughput m6 A sequencing ... The eluted RNA was treated with RNasin (Ambion Cat No. AM2694) according to the manufacturer’s instructions.

    Polyacrylamide Gel Electrophoresis:

    Article Title: RNA-Guided RNA Cleavage by a CRISPR RNA-Cas Protein Complex
    Article Snippet: Annealing was confirmed by non-denaturing 8% PAGE. .. In order to determine the divalent metal requirements of the purified complex, cleavage reactions were performed for 1 hour at 70°C in 50 mM HEPES pH 7.0, 250 mM KCl, 1 mM ATP, 10 mM DTT, 0.1 mM EDTA, and 1 mM metal (if applicable) in the presence of 1 unit of SUPERase-In ribonuclease inhibitor (Applied Biosystems).

    Lysis:

    Article Title: Enhanced Replication of Mouse Adenovirus Type 1 following Virus-Induced Degradation of Protein Kinase R (PKR)
    Article Snippet: .. Cells were collected at 24 hpi by scraping in ice-cold PBS containing 100 µg/ml cycloheximide, pelleting, and resuspending in 500 µl lysis buffer (20 mM Tris-Cl, 150 mM NaCl, 15 mM MgCl2 , 8% glycerol, 20 IU/ml SUPERase•In [Thermo Fisher Scientific catalog no. AM2696], 80 IU/ml murine RNase inhibitor [New England BioLabs catalog no. M0314S], 0.1 mg/ml heparin [Sigma H3393-50], 0.1 mg/ml cycloheximide, 1 mM DTT, 1× protease inhibitor [protease inhibitor cocktail kit; Thermo Scientific catalog no. 78410], 20 IU/ml Turbo DNase [Thermo Fisher Scientific catalog no. AM2238], 1% Triton X-100 [Sigma T9284]). .. Cells were lysed by passage through a chilled 26-gauge needle 10 times, vortex mixing for 30 s, and then incubating on ice for 5 min. Lysates were cleared by centrifugation for 5 min at 14,000 × g at 4°C.

    Article Title: KSHV induces immunoglobulin rearrangements in mature B lymphocytes
    Article Snippet: .. Single cell RT-PCR for immunoglobulin light chains Single cells were harvested by flow sorting into 96-well PCR plates containing 4μl of RNA lysis buffer (0.5x PBS+10mM DTT+4U SUPERas-In (Thermo Cat #AM2694)). .. Plates were sealed and stored at -80°C. cDNA was synthesized directly in wells using Thermo High Capacity cDNA synthesis kit (Cat #4368814).

    Article Title: FEM1 proteins are ancient regulators of SLBP degradation
    Article Snippet: Cell lysis was performed with lysis buffer (50 mM Tris, at pH 8.0, 150 mM NaCl, glycerol 10%, 1mM EDTA, 50 mM NaF, and NP-40 0.5%) supplemented with protease and phosphatase inhibitors. .. SUPERase-In™ RNase Inhibitor (Thermo Fisher Scientific) was used at 1U/μL where indicated.

    Article Title: KSHV induces immunoglobulin rearrangements in mature B lymphocytes
    Article Snippet: .. Single cells were harvested by flow sorting into 96-well PCR plates containing 4μl of RNA lysis buffer (0.5x PBS+10mM DTT+4U SUPERas-In (Thermo Cat #AM2694)). .. Plates were sealed and stored at -80°C. cDNA was synthesized directly in wells using Thermo High Capacity cDNA synthesis kit (Cat #4368814).

    Article Title: Accurate, Streamlined Analysis of mRNA Translation by Sucrose Gradient Fractionation
    Article Snippet: Lysis buffer (see Recipes) 10% sucrose solution (see Recipes) 50% sucrose solution (see Recipes) .. Reagents Cells-to-Ct kit (Thermo Fisher Scientific, Invitrogen™, catalog number: AM1728) Superase-In (Thermo Fisher Scientific, Invitrogen™, catalog number: AM2696) XenoRNA (Thermo Fisher Scientific, Invitrogen™, catalog number: 4386995, part of control kit) Note: Store in small aliquots at −80 °C.

    Agarose Gel Electrophoresis:

    Article Title: FEM1 proteins are ancient regulators of SLBP degradation
    Article Snippet: SUPERase-In™ RNase Inhibitor (Thermo Fisher Scientific) was used at 1U/μL where indicated. .. Lysates were immunoprecipitated with either Strep-Tactin (IBA) Superflow resin or anti-FLAG M2 affinity agarose gel (Sigma).

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Diverse motif ensembles specify non-redundant DNA binding activities of AP-1 family members in macrophages
    Article Snippet: .. First strand synthesis: We mixed fragmented RNA with 0.5 μL random primer (3 μg/μL) Life Tech #48190-011, 0.5 μL oligo-dT (50 μM from SSIII kit), 1 μL dNTPs (10 mM Life Tech, cat 18427088) and 0.5 μL SUPERase-In (ThermoFisher CatAM2696) and heat 50 °C for 1 min. .. We then added 5.8 μL ddH2 O, 0.1 μL actinomycin (2 μg/μL Sigma cat#A1410), 1 μL DTT (100 mM Life Tech cat# P2325), 0.2 μL of 1% Tween and 0.5 μL of Superscript III and incubate 25 °C for 10 min, then 50 °C for 50 min. Bead clean up: We added 36 μL of RNAClean XP (Ampure XP) and mixed, incubating for 15 min on ice.

    Article Title: Translation Enhancing ACA Motifs and Their Silencing by a Bacterial Small Regulatory RNA
    Article Snippet: The hybrid genes carried yifK wt or mutant 5′ UTR sequences fused to the cat -3×FLAG coding sequence (chloramphenicol acetyl transferase in-frame fusion to the 3×FLAG epitope). .. In addition to kit solutions A and B, reaction mix contained, 10 U of RNase inhibitor SUPERase (Ambion) and template plasmid DNA added to either 0.5 or 5 pM final concentration.

    Article Title: Desmoplasia and oncogene driven acinar-to-ductal metaplasia are concurrent events during acinar cell-derived pancreatic cancer initiation in young adult mice
    Article Snippet: RNA isolation RNA was extracted from microdissected specimens and KP pancreata using the PicoPure RNA Isolation Kit (Thermo Fisher, Cat# KIT0204) following the manufacturer’s instructions, including the optional on column DNAse digestion using RNase Free DNase (Qiagen, Cat79254). .. RNA was eluted using 11 μL of RNase Free ddH2 O and 0.5 μL of SUPERase•In RNase Inhibitor (Thermo Fisher, Cat# AM2694) was immediately added to eluted RNA.

    Chromatin Immunoprecipitation:

    Article Title: Desmoplasia and oncogene driven acinar-to-ductal metaplasia are concurrent events during acinar cell-derived pancreatic cancer initiation in young adult mice
    Article Snippet: RNA was eluted using 11 μL of RNase Free ddH2 O and 0.5 μL of SUPERase•In RNase Inhibitor (Thermo Fisher, Cat# AM2694) was immediately added to eluted RNA. .. RNA quality was quantified using the Agilent RNA 6000 Pico Chip (Agilent, Cat# 5067–1513) and Agilent 2100 Bioanalyzer platform.

    Plasmid Preparation:

    Article Title: Translation Enhancing ACA Motifs and Their Silencing by a Bacterial Small Regulatory RNA
    Article Snippet: .. In addition to kit solutions A and B, reaction mix contained, 10 U of RNase inhibitor SUPERase (Ambion) and template plasmid DNA added to either 0.5 or 5 pM final concentration. ..

    Sample Prep:

    Article Title: Unique Features of the m6A Methylome in Arabidopsis thaliana
    Article Snippet: The eluted RNA was treated with RNasin (Ambion Cat No. AM2694) according to the manufacturer’s instructions. .. TruSeq Stranded mRNA Sample Prep Kit (Illumina) was used to construct the library from immunoprecipitated RNA and input RNA according to a published protocol .

    In Vitro:

    Article Title: Translation Enhancing ACA Motifs and Their Silencing by a Bacterial Small Regulatory RNA
    Article Snippet: Paragraph title: In vitro translation ... In addition to kit solutions A and B, reaction mix contained, 10 U of RNase inhibitor SUPERase (Ambion) and template plasmid DNA added to either 0.5 or 5 pM final concentration.

    Immunoprecipitation:

    Article Title: FEM1 proteins are ancient regulators of SLBP degradation
    Article Snippet: SUPERase-In™ RNase Inhibitor (Thermo Fisher Scientific) was used at 1U/μL where indicated. .. Lysates were immunoprecipitated with either Strep-Tactin (IBA) Superflow resin or anti-FLAG M2 affinity agarose gel (Sigma).

    Article Title: Unique Features of the m6A Methylome in Arabidopsis thaliana
    Article Snippet: The eluted RNA was treated with RNasin (Ambion Cat No. AM2694) according to the manufacturer’s instructions. .. TruSeq Stranded mRNA Sample Prep Kit (Illumina) was used to construct the library from immunoprecipitated RNA and input RNA according to a published protocol .

    Stripping Membranes:

    Article Title: Diverse motif ensembles specify non-redundant DNA binding activities of AP-1 family members in macrophages
    Article Snippet: Collect beads on magnet and transfer eluate containing fragmented mRNA to a new PCR strip. .. First strand synthesis: We mixed fragmented RNA with 0.5 μL random primer (3 μg/μL) Life Tech #48190-011, 0.5 μL oligo-dT (50 μM from SSIII kit), 1 μL dNTPs (10 mM Life Tech, cat 18427088) and 0.5 μL SUPERase-In (ThermoFisher Cat#AM2696) and heat 50 °C for 1 min.

    Fractionation:

    Article Title: Enhanced Replication of Mouse Adenovirus Type 1 following Virus-Induced Degradation of Protein Kinase R (PKR)
    Article Snippet: Cells were collected at 24 hpi by scraping in ice-cold PBS containing 100 µg/ml cycloheximide, pelleting, and resuspending in 500 µl lysis buffer (20 mM Tris-Cl, 150 mM NaCl, 15 mM MgCl2 , 8% glycerol, 20 IU/ml SUPERase•In [Thermo Fisher Scientific catalog no. AM2696], 80 IU/ml murine RNase inhibitor [New England BioLabs catalog no. M0314S], 0.1 mg/ml heparin [Sigma H3393-50], 0.1 mg/ml cycloheximide, 1 mM DTT, 1× protease inhibitor [protease inhibitor cocktail kit; Thermo Scientific catalog no. 78410], 20 IU/ml Turbo DNase [Thermo Fisher Scientific catalog no. AM2238], 1% Triton X-100 [Sigma T9284]). .. After centrifugation, gradients were pumped out of the top with a Brandel BR-188 density gradient fractionation system with a continuous reading of the OD254 .

    High Throughput Screening Assay:

    Article Title: Unique Features of the m6A Methylome in Arabidopsis thaliana
    Article Snippet: Paragraph title: High-throughput m6 A sequencing ... The eluted RNA was treated with RNasin (Ambion Cat No. AM2694) according to the manufacturer’s instructions.

    Staining:

    Article Title: Selective ribosome profiling as a tool to study the interaction of chaperones and targeting factors with nascent polypeptide chains and ribosomes
    Article Snippet: .. 1-Ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride (EDC, Thermo Scientific, 22980) Glycine (Roth, 3790) NaHCO3 (Roth, 6885) Superase•In RNase inhibitor (Ambion, AM2696) Micrococcal nuclease (Nuclease S7) (Roche, 10107921001 or ) Ethylene glycol tetraacetic acid (EGTA, AppliChem, A0878) Sucrose (Sigma, 16104) Strep-Tactin Sepharose 50% suspension (IBA, 2-1201-025) DNA, sodium salt from salmon testes (Sigma-Aldrich, D1626) Kanamycin (Applichem, A1493) Isopropyl β-D-1-thiogalactopyranoside (IPTG, Roth, CN08) Colloidal coomassie staining solution (Roth, Roti-Blue quick, 4829) Imidazole (Roth, 3899) CAUTION Imidazole is corrosive. ..

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    Thermo Fisher 11836170001 superasin thermo am2696 rnasin promega n2515
    11836170001 Superasin Thermo Am2696 Rnasin Promega N2515, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 2 article reviews
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